FIGURE 1.

The G_q signaling and Ca²⁺ elicited SFK phosphorylation in platelets. A, washed platelets from wild-type (P2Y₁₂^+/+) and P2Y₁₂ deficient (P2Y₁₂^−/−) mice were preincubated with Ro-31-8220 (RO) (5 μm) or dimethyl sulfoxide (DMSO) for 5 min and then stimulated with AYPGKF (500 μm) at 37 °C for 5 min with stirring. SFK phosphorylation was detected by Western blotting with a rabbit monoclonal antibody specifically recognizing the phosphorylated Src residue Tyr⁴¹⁶. B, washed platelets from wild-type (G_q^+/+) and G_q deficient (G_q^−/−) mice were preincubated with Me₂SO, Cangrelor (1 μm), Ro-31-8220, or Cangrelor plus Ro-31-8220 for 5 min, and then stimulated with AYPGKF (500 μm) at 37 °C for 5 min with stirring. C, washed platelets from C57B6 mice were preincubated with Me₂SO, Cangrelor, or Ro-31-8220 for 5 min and then stimulated with serotonin (10 μm) at 37 °C for 5 min with stirring. D, washed platelets from TP deficient mice were stimulated with serotonin at 37 °C for 5 min with stirring. E, washed platelets from wild-type (G_q^+/+) and G_q deficient (G_q^−/−) mice were stimulated with A23187 (0.5 μm) at 37 °C for 5 min with stirring. F, washed platelets from TP deficient mice were preincubated with buffer, MRS2179 (MRS) (10 μm), Cangrelor, or MRS2179 plus Cangrelor for 5 min and then incubated with A23187 (0.5 μm) or buffer at 37 °C for 5 min with stirring.